Development of Specific Sequence-Characterized Amplified Region Markers for Detecting Histoplasma capsulatum in Clinical and Environmental Samples

Frías-De-León, María Guadalupe; López, Gabina Arenas; Taylor, Mária Lucía; Altamirano, Gustavo Acosta; Reyes-Montes, María del Roció

doi:10.1128/jcm.05271-11

Cited by 23 publications

(24 citation statements)

References 42 publications

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“…Recently, a comparison of different protocols for detection of H. capsulatum DNA by PCR showed that the sensitivity of the assay depended on both the selected target region and the type of amplification assay (conventional or rt-PCR) (24). Herein, the most sensitive and specific protocols were based on the amplification of the internal transcribed spacer (ITS) by rt-PCR (30), and the less sensitive protocols were based on the amplification of monocopy genes (Hc100p and SCAR 220 ) by conventional PCR (31,32).…”

mentioning

confidence: 99%

A Multiplex Real-Time PCR Assay for Identification of Pneumocystis jirovecii, Histoplasma capsulatum, and Cryptococcus neoformans/Cryptococcus gattii in Samples from AIDS Patients with Opportunistic Pneumonia

et al. 2014

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A molecular diagnostic technique based on real-time PCR was developed for the simultaneous detection of three of the most frequent causative agents of fungal opportunistic pneumonia in AIDS patients: Pneumocystis jirovecii, Histoplasma capsulatum, and Cryptococcus neoformans/Cryptococcus gattii. This technique was tested in cultured strains and in clinical samples from HIV-positive patients. The methodology used involved species-specific molecular beacon probes targeted to the internal transcribed spacer regions of the rDNA. An internal control was also included in each assay. The multiplex real-time PCR assay was tested in 24 clinical strains and 43 clinical samples from AIDS patients with proven fungal infection. The technique developed showed high reproducibility (r 2 of >0.98) and specificity (100%). For H. capsulatum and Cryptococcus spp., the detection limits of the method were 20 and 2 fg of genomic DNA/20 l reaction mixture, respectively, while for P. jirovecii the detection limit was 2.92 log 10 copies/20 l reaction mixture. The sensitivity in vitro was 100% for clinical strains and 90.7% for clinical samples. The assay was positive for 92.5% of the patients. For one of the patients with proven histoplasmosis, P. jirovecii was also detected in a bronchoalveolar lavage sample. No PCR inhibition was detected. This multiplex real-time PCR technique is fast, sensitive, and specific and may have clinical applications.

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confidence: 99%

A Multiplex Real-Time PCR Assay for Identification of Pneumocystis jirovecii, Histoplasma capsulatum, and Cryptococcus neoformans/Cryptococcus gattii in Samples from AIDS Patients with Opportunistic Pneumonia

et al. 2014

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“…However, none of these methods produced an amplicon from the 39 samples shown to be positive with the Hc100 nested PCR. Also, given that the trees obtained with the 3 different had the same topology, the classical genes proposed by Kasuga et al are adequate to achieve the genetic comparison inside the H. capsulatum population (34, 35, 37–39, 60).…”

Section: Discussionmentioning

confidence: 99%

“…Here we showed the presence of H. capsulatum in the Colombian environment and describe the genetic analysis of H. capsulatum Colombian population by MLST using both clinically relevant molecular markers as well as the sequences of genes used for diagnosis purposes including the partial sequence of 100 KDa protein (Hc100), M antigen (AgM) and two Sequences Characteristic Amplified Randomly (SCAR) (34–39).…”

Section: Introductionmentioning

confidence: 99%

Molecular epidemiology of ColombianHistoplasma capsulatumisolates shows their polyphyletic behavior and point out raw chicken manure as one of the infections sources

Gómez-Londoño

Arango-Arteaga

McEwen-Ochoa

et al. 2018

Preprint

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“…Most studies in the literature evaluated the performance of nested PCR in the diagnosis of DH, a method that is associated with the increased risk of amplicons contamination, since it involves the manipulation of amplified products. Many of the PCR tests for histoplasmosis were developed in-house [11][12][13][16][17][18][19][20][21][22][23][24][25][26][27]. The gene encoding a 100 kDa molecular protein, which is described as necessary for the survival of H. capsulatum in human cells, has been the most common target for the diagnosis of histoplasmosis [11,12,16,19,23,24,27].…”

Section: Overview Of the Performance Of Molecular Methods In The Diagmentioning

confidence: 99%

The Role of Molecular Tests in the Diagnosis of Disseminated Histoplasmosis

Vasconcellos

Lana

Pasqualotto

2019

JoF

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Histoplasmosis is an emerging fungal disease, with global distribution. The disseminated form of the disease is a more severe infection, generally associated with AIDS. Classic diagnostic methods for histoplasmosis consist of microscopy, culture, and histopathology. More recently, the importance of Histoplasma antigen detection has dominated the literature on histoplasmosis diagnosis, but the relevance of molecular assays has not been as much studied. Here we describe the results of a systematic literature review focusing on studies that mainly compared immunological techniques (Histoplasma urine antigen detection) with molecular tests for the diagnosis of histoplasmosis. In addition to the review of comparative studies using such diagnostic techniques, the literature on polymerase chain reaction (PCR) tests in patients with disseminated histoplasmosis is also summarized. Two studies reported the comparison between immunological and molecular methods applied simultaneously for the diagnosis of disseminated histoplasmosis. PCR demonstrates a satisfactory performance assisting in the detection of Histoplasma spp. DNA in clinical samples.

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Development of Specific Sequence-Characterized Amplified Region Markers for Detecting Histoplasma capsulatum in Clinical and Environmental Samples

Cited by 23 publications

References 42 publications

A Multiplex Real-Time PCR Assay for Identification of Pneumocystis jirovecii, Histoplasma capsulatum, and Cryptococcus neoformans/Cryptococcus gattii in Samples from AIDS Patients with Opportunistic Pneumonia

A Multiplex Real-Time PCR Assay for Identification of Pneumocystis jirovecii, Histoplasma capsulatum, and Cryptococcus neoformans/Cryptococcus gattii in Samples from AIDS Patients with Opportunistic Pneumonia

Molecular epidemiology of ColombianHistoplasma capsulatumisolates shows their polyphyletic behavior and point out raw chicken manure as one of the infections sources

The Role of Molecular Tests in the Diagnosis of Disseminated Histoplasmosis

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